Histomorphological Changes Research Articles

Morin Improves the Bone Histomorphology and Biochemical Markers in an Animal Model of Ovariectomy‐Induced Osteoporosis by Suppressing Autophagy and Apoptosis

ABSTRACTOsteoporosis (OP) is the most prevalent metabolic bone disease and an important postmenopausal consequence. This study aimed to investigate the effects of morin, a flavonoid with beneficial properties, on ovariectomy‐induced OP. Animals were ovariectomized (OVX) and treated with different doses of morin (15, 30, and 45 mg/kg/day) or estradiol (10 μg/kg/day) for 10 weeks by gavage. Then bone histo‐stereology, bone‐related biochemical indicators, and gene and protein levels of autophagy and apoptosis‐related markers were analyzed. In comparison to controls, OVX significantly decreased the number of osteoblasts (5.78 × 106 vs. 1.66 × 106) and osteocytes (32.55 × 106 vs. 11.92 × 106), whereas increasing the number of osteoclasts (83.38 × 103 vs. 392.1 × 103). Moreover, OVX caused a remarkable decrease in bone structures and Ca, P, and estradiol levels while increasing ALP and OC (p < 0.001). The administration of 45 mg/kg/day morin restored the effects of OP on bone histomorphology and biochemical markers (p < 0.05). Further studies revealed that morin caused a 7.1% and 36.6% decrease in the bone level of LC3 and BECN1 proteins, respectively, compared to the OVX group. Also, morin caused a significant decrease of 47.4% in the CASP3 level and a significant increase of 23.6% in the BCL‐2 level compared to OVX animals (p < 0.001). The present findings showed that morin is potentially able to improve the bone‐related histomorphological and biochemical changes caused by osteoporosis, which is probably attributed to the suppression of apoptosis‐ and autophagy‐caused cell death.

A preliminary study on the effects of Xiang Shao granules on reproductive endocrinology in drugged ovariectomised rats

ObjectiveTo establish a rat model of pharmacological ovariectomy by GnRH-a injection and to preliminarily investigate the reproductive endocrine effects of Xiangshao granules on pharmacologically ovariectomized rats. Methods: A rat model of pharmacological ovariectomy was established by injecting female rats with Gonadotropin-releasing hormone agonist(GnRH-a).The rats were randomly divided into four groups: GnRH-a injected saline group (GnRH-a + NS); GnRH-a injected oestradiol group (GnRH-a + E2); GnRH-a injected Xiangshao granule group (GnRH-a + Xiangshao), and the control group of saline-injected rats (NS + NS). The number of rats per group was 6.According to observations of the rats' vaginal smears, modelling was determined as successful. Then corresponding drug gavage intervention was administered for 28 days, and rat body weight and anal temperature were measured every other day to adjust the drug intervention amount according to body weight changes. Plasma sex hormone levels (E2, FSH, LH), uterine weight, uterine index and endometrial histomorphological changes, ovarian weight, and ovarian index and ovarian histomorphological changes were measured in each group after the gavage.Results(1) Plasma sex hormone levels (E2, FSH, LH) of the GnRH-a + NS, GnRH-a + E2, and GnRH-a + Xiangshao granule groups were significantly lower than the NS + NS group (P < 0.001), while the E2 level of the GnRH-a + E2 group was higher than that of the GnRH-a + Xiangshao granule group (P < 0.05). The FSH level of the GnRH-a + E2 group was significantly lower than that of the GnRH-a + Xiangshao granule group (P < 0.05). The LH level of the GnRH-a + E2 group was significantly lower than those in the GnRH-a + NS and GnRH-a + Xiangshao granule groups (P < 0.001, P = 0.001). The LH and FSH levels of the GnRH-a + NS and GnRH-a + Xiangshao granule groups were not significantly different (P > 0.05). (2) Compared with the NS + NS group, the uterine weight and uterine index, and ovarian weight and ovarian index of GnRH-a injected rats in each model all significantly decreased (P < 0.001). Between the groups, the uterine weight and uterine index, and ovarian weight and ovarian index of GnRH-a + E2 and GnRH-a + Xiangshao granule groups were all significantly higher than those of the GnRH-a + NS group (P < 0.001, P < 0.05). The uterine weight and uterine index, and ovarian weight and ovarian index of the GnRH-a + E2 group increased compared with the GnRH-a + Xiangshao granule group (P < 0.05). (3) Compared with the NS + NS group, the number of primordial follicles of the GnRH-a + NS, GnRH-a + E2, and GnRH-a + Xiangshao granule groups increased significantly and the number of growing follicles and mature follicles significantly decreased. (4) Rats' uterine wall of the NS + NS and various GnRH-a groups was significantly thinner, with the endothelial layer atrophied, while the uterine wall of the GnRH-a + E2 and GnRH-a + Xiangshao granule groups was thicker obviously, with the number of vaginal folds and blood vessels also increasing. Specifically, the uterus and vagina improvements in the GnRH-a + E2 group were more obvious than in GnRH-a + NS and GnRH-a + Xiangshao granule groups.ConclusionGnRH-a injection can reduce the levels of sex hormones E2, FSH, and LH in rats, causing perimenopausal symptoms such as hot flashes, while Xiangshao and E2 granules could significantly improve such symptoms and exert a slight oestrogenic effect, to a lesser extent than E2 does.Trial registrationNot applicable.

Targeting the Sirtuin–1/PPAR–Gamma Axis, RAGE/HMGB1/NF-κB Signaling, and the Mitochondrial Functions by Canagliflozin Augments the Protective Effects of Levodopa/Carbidopa in Rotenone-Induced Parkinson’s Disease

Background and Objectives: Parkinson’s disease (PD) is a pathological state characterized by a combined set of abnormal movements including slow motion, resting tremors, profound stiffness of skeletal muscles, or obvious abnormalities in posture and gait, together with significant behavioral changes. Until now, no single therapeutic modality was able to provide a complete cure for PD. This work was a trial to assess the immunomodulatory effects of canagliflozin with or without levodopa/carbidopa on rotenone-induced parkinsonism in Balb/c mice. Materials and Methods: In a mouse model of PD, the effect of canagliflozin with or without levodopa/carbidopa was assessed at the behavioral, biochemical, and histopathological levels. Results: The combination of levodopa/carbidopa and canagliflozin significantly mitigated the changes induced by rotenone administration regarding the behavioral tests, striatal dopamine, antioxidant status, Nrf2 content, SIRT–1/PPAR–gamma axis, RAGE/HMGB1/NF-κB signaling, and mitochondrial dysfunction; abrogated the neuroinflammatory responses, and alleviated the histomorphologic changes induced by rotenone administration relative to the groups that received either levodopa/carbidopa or canagliflozin alone. Conclusions: Canagliflozin may represent a new adjuvant therapeutic agent that may add value to the combatting effects of levodopa/carbidopa against the pathological effects of PD.

Protective Effects of Astaxanthin against Oxidative Stress: Attenuation of TNF-α-Induced Oxidative Damage in SW480 Cells and Azoxymethane/Dextran Sulfate Sodium-Induced Colitis-Associated Cancer in C57BL/6 Mice.

In this study, we investigated the protective effects of astaxanthin (AST) against oxidative stress induced by the combination of azoxymethane (AOM) and dextran sulfate sodium (DSS) in colitis-associated cancer (CAC) and TNF-α-induced human colorectal cancer cells (SW480), as well as the underlying mechanism. In vitro experiments revealed that astaxanthin reduced reactive oxygen species (ROS) generation and inhibited the expression of Phosphorylated JNK (P-JNK), Phosphorylated ERK (P-ERK), Phosphorylated p65 (P-p65), and the NF-κB downstream protein cyclooxygenase-2 (COX-2). In vivo experiments showed that astaxanthin ameliorated AOM/DSS-induced weight loss, shortened the colon length, and caused histomorphological changes. In addition, astaxanthin suppressed cellular inflammation by modulating the MAPK and NF-κB pathways and inhibiting the expression of the proinflammatory cytokines IL-6, IL-1β, and TNF-α. In conclusion, astaxanthin attenuates cellular inflammation and CAC through its antioxidant effects.

To Evaluate The Histomorphological Changes Of Postmortem Transthoracic Needle Biopsy/Autopsy Lung In COVID-19 Patients

Objective: To study the histomorphological changes in postmortem needle biopsy/autopsy lung in COVID-19 Patients. Study Design: This was an observational study done at Shri Atal Bihari Vajpayee Medical College &amp; Research Institute, Bengaluru, and Karnataka from March 2021 to May 2021. Methodology: A total of 30 patients who died of Covid-19 were included and all of them were diagnosed to be Covid -19 positive by RT-PCR testing. Post-mortem transthoracic needle biopsy was done to obtain lung tissue by using B-Bard biopsy gun in midaxillary approach. The biopsy samples were grossed and processed after fixing for 48hrs. The slides obtained were stained by routine hematoxylin and eosin stain, studied for morphology of the lung tissue and findings were recorded. Results: We obtained consent for post-mortem transthoracic needle biopsy/autopsy of lung in 30 deceased cases. All these cases were positive for SARS –COV2 on RT-PCR. The age of the cases was ranging from 26-72 years and median age is 53years, males (63.3%) were affected more than females (36.7%). Most deceased had comorbidities, among them diabetes mellitus (26.6%) and hypertension (26.6%) were most common. Lung biopsy samples were taken from 30 deceased patients. Histomorphological feature of diffuse alveolar damage was seen in 90% (27 out of 30) cases. Other findings noted on microscopy were chronic interstitial inflammation in 6.7% cases, pulmonary edema (3.33%), intra alveolar haemorrhage (3.33%) and viral cytopathic changes (3.33%). Conclusion: This study highlighted the pulmonary morphological changes which are most prominent in COVID-19 infections, but it was a difficult to ascertain that the findings are related to viral infection or due to associated coomorbidities

The neural regulation and impact of posterior nasal neurectomy on nasal ciliary motion in vivo in a murine allergic rhinitis model.

Posterior nasal neurectomy (PNN) is a commonly employed surgical approach for the treatment of allergic rhinitis (AR). Due to its denervation effect on the nasal mucosa, PNN may potentially alter the motion and defensive capability of cilia. Previous research on the effects of neural regulation and denervation on cilia has been limited by the absence of a feasible in vivo evaluation method for assessing ciliary function. Utilizing a new system developed by our team for visualizing and analyzing ciliary motion in vivo, we analysed ciliary beat frequency and distance in vivo and histomorphological changes in a murine PNN and AR model. Ovalbumin, histamine and neurotransmitters (acetylcholine chloride, α receptor agonist and β receptor agonist) were applied to investigate the responsiveness and neural regulation of the nasal mucosa. Denervation resulting from PNN led to a reduction in nasal ciliary beat frequency (CBF) to 78% of the control, as well as diminished response towards allergens and histamine. Among neurotransmitters examined, α receptor agonists exhibited inhibitory effects on in vivo ciliary motion while acetylcholine and β receptor agonists demonstrated stimulatory effects. PNN did not affect the reactivity of in vivo cilia towards these neurotransmitters. PNN-induced denervation can reduce ciliary motion, potentially compromising the defensive capability of nasal mucosa. Neural regulation and the neurotransmitters involved have significant effect on ciliary motion.

Histomorphological Changes in Testis of Rats with Prolonged Exposure to Allethrin-Based Mosquito Coil Smoke.

To explore the potential adverse effects of prolonged inhalation of mosquito coil smoke on the testicular histomorphology and serum testosterone levels in rats. An experimental study. Place and Duration of the Study: Department of Anatomy, Army Medical College, National University of Medical Sciences (NUMS), Rawalpindi, Pakistan, from January to December 2020. This study was carried out on 20 male Sprague-Dawley rats, divided into control Group A (n = 10) and experimental Group B (n = 10). Rats in Group B were exposed to mosquito coil smoke (allethrin-based) for 4 hours / day for 12 weeks. After 12 weeks of exposure, serum testosterone levels and testicular morphology (seminiferous tubule diameter, germinal epithelium height, and testicular capsule thickness) were compared between the groups. Rats exposed to mosquito coil smoke showed significantly reduced serum testosterone levels (p <0.001) along with testicular histological disruption in terms of significantly dilated seminiferous tubules (p <0.001), reduced germinal epithelial height (p <0.001), and thickened testicular capsule (p <0.001), as compared to the control group. Prolonged inhalation of allethrin-based mosquito coil smoke reduced serum testosterone levels and caused testicular histological disruption in the exposed group of rats. Allethrin, Germinal epithelium, Mosquito coil, Seminiferous tubules, Testicular capsule, Testosterone, Testis.

Placental growth factor alleviates hyperglycemia-induced trophoblast pyroptosis by regulating mitophagy.

Hyperglycemia is closely related to trophoblast dysfunction during pregnancy and results in suppressed invasion, migration, and pro-inflammatory cell death of trophoblasts. Hyperglycemia is a dependent risk factor for gestational hypertension accompanied by decreased placental growth factor (PLGF), which is important for maternal and fetal development. However, there is currently a lack of evidence to support whether PLGF can alleviate trophoblast cell dysfunction caused by high blood sugar. Here, we aim to clarify the effect of hyperglycemia on trophoblast dysfunction and determine how PLGF affects this process. The changes in placental tissue histomorphology from gestational diabetes mellitus (GDM) patients were compared with those of normal placentas. HTR8/SVneo cells were cultured in different amounts of glucose to examine cellular pyroptosis, migration, and invasion as well as PLGF levels. Furthermore, the levels of pyroptosis-related proteins (NLRP3, pro-caspase1, caspase1, IL-1β, and Gasdermin D [GSDMD]) as well as autophagy-related proteins (LC3-II, Beclin1, and p62) were examined by Western blotting. The GFP-mRFP-LC3-II system and transmission electron microscopy were used to detect mitophagy levels, and small interfering RNAs targeting BCL2 Interacting Protein 3 (siBNIP3) and PTEN-induced kinase 1 (siPINK1) were used to determine the role of mitophagy in pyroptotic death of HTR-8/SVneo cells. Our results show that hyperglycemia upregulates NLRP3, pro-caspase1, caspase1, IL-1β at the protein level in GDM patients. High glucose (HG, 25 mM) inhibits viability, invasion, and migration of trophoblast cells while suppressing superoxide dismutase levels and promoting malondialdehyde production, thus leading to a senescence associated beta-gal-positive cell burst. PLGF levels in nucleus and the cytosol are also inhibited by HG, whereas PLGF treatment inhibited pyroptosis-related protein levels of NLRP3, pro-caspase1, caspase1, IL-1β, and GSDMD, Gasdermin D N-terminal domain (GSDMD-N). HG-induced mitochondrial dysfunction and BNIP3 and PINK1/Parkin expression. Knocking down BINP3 and PINK1 abolished the protective role of PLGF by preventing mitophagy. PLGF inhibited hyperglycemia, while PLGF reversed hyperglycemic injury by promoting mitophagy via the BNIP3/PINK1/Parkin pathway. Altogether, these results suggest that PLGF may protect against trophoblast dysfunction in diabetes.

Neuroprotective effects of ulinastatin on Escherichia coli meningitis rats through inhibiting PKCα phosphorylation and reducing zonula occludens-1 degradation

Ulinastatin, a broad-spectrum inflammatory inhibitor widely employed in the management of severe pancreatitis and sepsis, has not been extensively investigated for its therapeutic potential in bacterial meningitis. This study aims to assess the neuroprotective effects of ulinastatin on bacterial meningitis and elucidate its underlying mechanism. The rat model of bacterial meningitis was established by intracerebral injection of Escherichia coli. 3-week-old SD rats were randomly divided into 5 groups with 8 rats in each group, including control group, E.coli group, E.coli + UTI group (ulinastatin 50000IU/kg), E.coli + UTI + PMA group (ulinastatin 50000IU/kg + PMA 200 ug/kg), and E.coli + PMA group(PMA 200 ug/kg). Behavioral changes were assessed by Loeffler neurobehavioral score. Histomorphologic changes and apoptosis were assessed by hematoxylin and eosin staining, Nissl staining and TUNEL staining. Immunohistochemistry and immunofluorescence and western blotting were used to detect the expression levels of zonula occludens-1 (ZO-1) and phosphorylation protein kinase C (PKCα).It was found that ulinastatin treatment in Escherichia coli meningitis rats improved neurological function, alleviated meningeal inflammatory infiltration, reduced neuronal death, promoted the integrity of the blood–brain barrier structure. Moreover, phorbol myristate acetate (PMA, a protein kinase C activator), blocked the effective action of ulinastatin. These findings suggest that ulinastatin had neuroprotective effects on bacterial meningitis by inhibiting PKCα phosphorylation and reducing ZO-1 degradation, demonstrating that ulinastatin may be a promising strategy in the treatment of bacterial meningitis.

Post-mortem Interval estimate based on dental pulp: A histomorphology approach.

Estimating the post-mortem interval (PMI) of human remains based on the histomorphology of dental pulp parameters is promising, but available evidence is scarce and sometimes contradictory without a scientific model. The aim of the study is to characterise the histomorphological changes of dental pulp associated with the decomposition of human remains by a qualitative and quantitative approach. The main aim is to establish a correlation based on post-mortem (PM) dental pulp histomorphology and the PMI, and whether pulp degradation could be an available medico-legal tool for PMI estimation beyond the first week after death (late PMI). The eligible sample consisted of 27 sound teeth from 16 healthy patients aged 16 to 72 years due to orthodontic or oral surgery treatment, to create PMI's simulating the death of the subject as the time elapsed from tooth avulsion. Data collected from patients (sex, date of birth, tooth position, date and hour of the avulsion, date and hour of pulp extraction) were anonymised in accordance with the requirements of Faculty of Dental Medicine of the University of Lisbon. The sample was divided into 9 groups of 3 teeth according to different PMI sets from T0 (baseline) up to 2 weeks (T0, 7, 12, 24, 36, 48, and 72 hours, 1 and 2 weeks). All the dental samples were stored at room temperature up to the time of pulp extraction and then prepared with haematoxylin and eosin stain. High-resolution microscopy was performed to obtain histological images. An operator performed the qualitative evaluation of blood vessels, collagen fibres, and the extra-cellular matrix (ECM) in PM pulps and measured the variation in cells/nuclei density by counting 6 different ROIs (Regions of Interest) for each pulp manually and automatically (quantitative analysis). Qualitative results showed that the degeneration of dental pulp appears 7 hours after death but histological changes in vessels, fibres, and ECM in PM dental pulp are characterised by high variability, consequently it is not possible to generalise the results for early PMIs. Quantitative measurements proved that cell count cannot be standardised due to the presence of superimposed layers of cells and nuclei fragmentation. Odontoblasts did not demonstrate evidence of cellular or nuclear lysis up to 14 PM suggesting their applicability in late PMIs. Future research will focus on late PMIs and different techniques of tooth preparation.

Histomorphological and Dynamical Changes in Female River Lampreys during Maturation under Controlled Conditions as a Part of Lamprey Restoration Programs.

More than 40 species of lampreys (Petromyzontiformes) are known worldwide. Some of them are parasitic and feed on the blood of fish or other aquatic animals. Lampreys spawn once in their lifetime, after which they die. One of the representatives of the ichthyofauna of European rivers is the river lamprey, Lampetra fluviatilis. The river lamprey is now an endangered species due to loss and degradation of their habitats. The present study investigated gonadal development without hormonal stimulation in female river lampreys during puberty under controlled conditions for a period of seven months. Female river lampreys were kept in conditions that mimicked the natural environment. During the November-May period, samples were taken monthly to determine the extent of gonadal development and gastrointestinal and liver changes using histological examination. From the results obtained, the dynamical changes were determined for the following: gonadosomatic index (GSI; %), hepatosomatic index (his; %), and digestivesomatic index (DSI; %). With the gonadal development of female lampreys, an increase in GSI (7-23%; November-May) and a decrease in DSI (0.4-0.1%; November-May) histological changes were observed in the gonads (oocyte development), intestines (over time, decreased lipid vacuoles and enterocyte apoptosis), and in the liver (decreased lipid vacuoles and hepatocyte apoptosis over time) and in the digestive system resulting from its degradation. Also, it was observed that there was a change in the color of the liver to green in April. This study demonstrated the key role of the liver in the oocyte maturation process in this species.

Spectrum of various histomorphological changes in heart studied with autopsy specimens - An institutional observational study

Spectrum of various histomorphological changes in heart studied with autopsy specimens - An institutional observational study

Inter-strain variability in responses to a single administration of the cannabidiol-rich cannabis extract in mice

Cannabidiol (CBD) has gained widespread popularity; however, its pharmacological and toxicological profiles in the context of human genetic diversity remain largely unexplored. Here, we investigated the variability in metabolism and toxicity of CBD-rich cannabis extract (CRCE) in genetically diverse mouse models: C57BL/6J, B6C3F1/J, and NZO/HlLtJ strains. Mice received a single dose of CRCE containing 57.9% CBD at dosages of 0, 246, 738, and 2460 mg/kg of CBD. At 24 h after treatment, no appreciable histomorphological changes were detected in the liver. Plasma bilirubin levels increased markedly in all strains at the highest CBD dose. Mice in all treatment groups displayed significant but distinct increases in ALT and AST levels. While B6C3F1/J and NZO/HlLtJ mice had negligible plasma CBD levels at 738 mg/kg, C57BL/6J mice exhibited levels exceeding 7000 ng/mL. At 2460 mg/kg, high CBD concentrations were found in B6C3F1/J and C57BL/6J mice, but markedly lower levels were seen in NZO/HlLtJ mice. Gene expression profiling showed significant increases in Cyp2b10 across all strains but varying responses in Cyp1a1 expression, indicating strain-specific CYP dysregulation. Genetically diverse mice exhibited differential pharmacological and toxicological responses to CRCE, suggesting a high potential for inter-individual variability in the pharmacology and toxicology of CBD in humans.

Evaluation of Respiratory Toxicity Potential of Acrylamide in Zebrafish: Oxidative Stress Parameters, mRNAs Expression, and Histomorphological Changes in Gills

Evaluation of Respiratory Toxicity Potential of Acrylamide in Zebrafish: Oxidative Stress Parameters, mRNAs Expression, and Histomorphological Changes in Gills

Qing Hua Chang Yin ameliorates chronic colitis in mice by inhibiting PERK–ATF4–CHOP pathway of ER stress and the NF-κB signalling pathway

Context Ulcerative colitis has been clinically treated with Qing Hua Chang Yin (QHCY), a traditional Chinese medicine formula. However, its precise mechanisms in mitigating chronic colitis are largely uncharted. Objective To elucidate the therapeutic efficiency of QHCY on chronic colitis and explore its underlying molecular mechanisms. Materials and methods A total ion chromatogram fingerprint of QHCY was analysed. Chronic colitis was induced in male C57BL/6 mice using 2% dextran sodium sulphate (DSS) over 49 days. Mice were divided into control, DSS, DSS + QHCY (0.8, 1.6 and 3.2 g/kg/d dose, respectively) and DSS + mesalazine (0.2 g/kg/d) groups (n = 6). Mice were intragastrically administered QHCY or mesalazine for 49 days. The changes of disease activity index (DAI), colon length, colon histomorphology and serum pro-inflammatory factors in mice were observed. RNA sequencing was utilized to identify the differentially expressed transcripts (DETs) in colonic tissues and the associated signalling pathways. The expression of endoplasmic reticulum (ER) stress-related protein and NF-κB signalling pathway-related proteins in colonic tissues was detected by immunohistochemistry staining. Results Forty-seven compounds were identified in QHCY. Compared with the DSS group, QHCY significantly improved symptoms of chronic colitis like DAI increase, weight loss, colon shortening and histological damage. It notably reduced serum levels of IL-6, IL-1β and TNF-α. QHCY suppressed the activation of PERK–ATF4–CHOP pathway of ER stress and NF-κB signalling pathways in colonic tissues. Discussion and conclusions The findings in this study provide novel insights into the potential of QHCY in treating chronic colitis patients.

A Histomorphological and Histochemical changes of Postnatal Developmental in the Ovaries and Uterus of Local Rabbits (Oryctolagus cuniculus) in Duhok Province

A Histomorphological and Histochemical changes of Postnatal Developmental in the Ovaries and Uterus of Local Rabbits (Oryctolagus cuniculus) in Duhok Province

Methionine supplementation regulates eggshell quality and uterine transcriptome in late-stage broiler breeders

This study aimed to compare the effects of dietary methionine (Met) and 2-hydroxy-4-(methylthio)-butanoate (HMTBA) on the eggshell quality of broiler breeder hens and elucidate the mechanism of Met in improving eggshell quality from the perspectives of eggshell microstructure and shell gland physiological function. A total of 720 WOD188 broiler breeder hens at 40 wk old were assigned to 3 groups, with 8 replicates per group and 30 birds per replicate. Over 7 weeks, birds were fed a basal diet or the same diet supplemented with 0.15% Met or 0.17% HMTBA. Our findings revealed significant improvements in the Met group for egg shape index, shell thickness, breaking strength, and fracture toughness (P < 0.05), whereas the HMTBA group showed no significant improvements (P > 0.05). Met supplementation increased calcium and phosphorus levels in both serum and shell gland tissue (P < 0.05), and enhanced Ca2+ ATPase activity in shell gland tissue (P < 0.05). Histomorphological changes included enhanced mucosal fold dimensions and increased epithelial height in the shell gland (P < 0.05). Met also improved eggshell ultrastructure, resulting in a thicker effective layer and broader mammillae with fewer type B structures (P < 0.05). The mRNA levels for genes regulating eggshell ultrastructure, such as OC-116, CALB1, and ITM2C, were significantly upregulated in the Met group (P < 0.05). Transcriptome analysis identified 248 differentially upregulated genes in the Met group, primarily linked to the non-canonical Wnt/Ca2+ signaling pathway, crucial for calcium ion transport and cellular proliferation. This research highlights that Met supplementation improves eggshell quality by enhancing calcium transport and cellular proliferation in uterine function, particularly through the modulation of WNT11 and CALB1, influencing calcium deposition and ultrastructural development.

DELAYED EFFECTS OF RADIATION: A LITERATURE REVIEW

Relevance: The effect of radiation on the immune system of the offspring of persons affected by exposure to low doses of various radiation types is still an urgent issue. In the literature, several issues related to delayed consequences, namely, morphofunctional disorders in the organs of the descendants of irradiated individuals, have not received adequate resolution. The purpose was to analyze literature on the long-term effects of radiation, manifested by morphofunctional disorders of the immunocompetent organs in the descendants of persons exposed to radiation, to confirm the importance of further study of this issue. Methods: The review included publications indexed in PubMed, Medline, E-library, and CyberLeninka databases. The search was made using the Google Scholar search engine. The applied search filters included conducted experimental studies over the past eight years (2015-2023). Results: The search revealed insufficient publications devoted to studying metabolic processes and histomorphological changes in the immunocompetent organs of the body of the descendants of exposed individuals. An insufficient number of publications on long-term biochemical and pathohistological changes in immune system organs due to irradiation necessitate a profound study of the topic. Conclusion: The study of morphofunctional disorders in the descendants of the first generation of irradiated individuals remains a task for radiobiologists and oncologists, with the development of criteria for morphological diagnosis.

Direct Immunofluorescence in Cutaneous Vesiculobullous Lesions: A Cross-sectional Study in a Tertiary Care Center

Introduction: Cutaneous noninfectious autoimmune vesiculobullous diseases are a wide variety of lesion. They can show clinical and histopathological overlap, posing diagnostic challenges. Clinically, vesiculobullous lesions might not present with typical morphology and distribution in all cases. The specific histomorphological changes are observed only in an early and intact bulla. Immunofluorescence (IF) is a histochemical staining technique which detects antibodies in the tissue and body fluids. IF an adjuvant to histopathology is considered as gold standard in the diagnosis of vesiculobullous diseases. Methodology: A two-year retrospective cross-sectional descriptive study was conducted in the department of pathology, in a tertiary care center from January 2018 to December 2019. Paraffin embedded hematoxylin and eosin slides were retrieved from the archives and analyzed. Biopsies received in Michel’s medium were utilized for immunofluorescence study. Results: A total of 67 cases with Vesiculobullous lesions were analysed. The most common lesion diagnosed was Bullous Pemphigoid (BP) (29 cases -46.7%), followed by Pemphigus Vulgaris (PV) (19 cases- 30.6%). The highest number of cases were noted in the 7th decade, with a slight female preponderance. Sixteen out of 19 cases of pemphigus vulgaris and 28 out of 29 cases of bullous pemphigoid showed positive histomorphological correlation with IF. The PPV for pemphigus vulgaris was 100% and sensitivity was 84.2%, and in bullous pemphigoid, both the PPV and sensitivity was 96.5%. Higher concordance was noted between Histopathology and IF diagnosis in cases of BP in comparison to PV. Conclusion: Vesiculobullous lesions are heterogeneous in nature and can have clinical and histopathological overlap, posing diagnostic challenges. IF is considered gold standard and is necessary for the diagnosis in every case. A combined clinical, histopathological and IF examination is the best approach to accurate diagnosis. Keywords Bullous, Pemphigus, Vesiculobullous, Direct Immunofluorescence

Fagopyrum Dibotrys Rhizoma regulates pulmonary lipid metabolic homeostasis and the ERK-cPLA2 pathway to alleviate asthma in mice

BackgroundAsthma is a complex disease with mechanisms involving multiple factors, and there is still a lack of highly effective and low-side-effect drugs. Traditional Chinese medicine Fagopyrum Dibotrys Rhizoma (FDR) has been applied for the treatment of acute and chronic bronchitis as well as bronchial asthma due to its favorable pharmacological activity. However, the exact mechanism of FDR remains unclear. ObjectiveA mouse model of asthma was created using OVA and HDM. To investigate the mechanism of FDR in asthma treatment, a combination of network pharmacology, lipidomics, and molecular biology approaches was employed. MethodsTo evaluate the therapeutic effects of FDR on asthma, we established two distinct models of asthma in C57BL/6 J mice using OVA and HDM, respectively. We then employed LC-MS to analyze the major chemical constituents in FDR. Next, the network pharmacology approach was used to predict the potential targets and mechanisms of FDR in asthma treatment. Additionally, lipidomics analysis of mouse serum was conducted using LC-MS. Finally, the impact of FDR on the ERK -cPLA2 signaling pathway was investigated through Western Blotting assay. ResultsFDR treatment has been shown to improve histomorphological changes, lung function and inflammation in models of OVA and HDM-induced asthma. Using UPLC/LTQ-Orbitrap-MS, we were able to identify 12 potential active components. Network pharmacology analysis revealed that FDR shares 75 targets with asthma. Further analysis using GO and KEGG pathways demonstrated the involvement of key pathways such as PI3K-Akt, TNF, and MAPK. Additionally, lipidomics analysis of the serum from OVA and HDM induced asthma mice showed disturbances in lipid metabolism, which were effectively ameliorated by FDR treatment. Mechanistically, FDR inhibits ERK1/2-cPLA2, leading to a reduction in lysophospholipids and restoration of lipid balance, thereby aiding in the treatment of asthma. ConclusionFDR has been shown to improve lipid metabolism disorder in the serum of asthmatic mice, thereby potentially serving as a treatment for asthma. This can be achieved by regulating the activation levels of ERK1/2 and p38MAPK. Consequently, the production of lysophosphatide is reduced, thereby alleviating the disorder of lipid metabolism and achieving the desired therapeutic effect in asthma treatment.